Antibodies that bind human metapneumovirus fusion protein and their use

Abstract

Antibodies and antigen binding fragments that specifically bind to human metapneumovirus (hMPV) F protein and neutralize hMPV are disclosed. Nucleic acids encoding these antibodies, vectors and host cells are also provided. The disclosed antibodies, antigen binding fragments, nucleic acids and vectors can be used, for example, to inhibit an hMPV infection or detect a hMPV infection.

We claim: 
     
         1 . An isolated monoclonal antibody or antigen binding fragment thereof, comprising:
 a) a heavy chain variable region (V H ) and a light chain variable region (V L ) comprising a heavy chain complementarity determining region (HCDR) 1, a HCDR2, and a HCDR3, and a light chain complementarity determining region (LCDR) 1, a LCDR2, and a LCDR3 of the V H  and V L  set forth as SEQ ID NOs: 1 and 2, respectively;   b) a heavy chain variable region and a light chain variable region comprising a HCDR1, a HCDR2, and a HCDR3, and a LCDR1, a LCDR2, and a LCDR3 of the V H  and V L  set forth as SEQ ID NOs: 11 and 12, respectively;   c) a heavy chain variable region and a light chain variable region comprising a HCDR1, a HCDR2, and a HCDR3, and a LCDR1, a LCDR2, and a LCDR3 of the V H  and V L  set forth as SEQ ID NOs: 21 and 22, respectively;   d) a heavy chain variable region and a light chain variable region comprising a HCDR1, a HCDR2, and a HCDR3, and a LCDR1, a LCDR2, and a LCDR3 of the V H  and V L  set forth as SEQ ID NOs: 31 and 32, respectively;   e) a heavy chain variable region and a light chain variable region comprising a HCDR1, a HCDR2, and a HCDR3, and a LCDR1, a LCDR2, and a LCDR3 of the V H  and V L  set forth as SEQ ID NOs: 49 and 50, respectively; or   f) a heavy chain variable region and a light chain variable region comprising a HCDR1, a HCDR2, and a HCDR3, and a LCDR1, a LCDR2, and a LCDR3 of the V H  and V L  set forth as SEQ ID NOs: 59 and 60, respectively,   wherein the monoclonal antibody specifically binds to human metapneumovirus (hMPV) F protein and neutralizes hMPV, and   wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 are identified using Kabat, Chothia or IMGT numbering.   
     
     
         2 . The isolated monoclonal antibody or antigen binding fragment of  claim 1 , wherein
 a) the HCDR1, the HCDR2, the HCDR3, the LCDR1, the LCDR2, and the LCDR3 comprise the amino acids sequences set forth as SEQ ID NOs: 5, 6, 7, 8, 9 and 10, respectively;   b) the HCDR1, the HCDR2, the HCDR3, the LCDR1, the LCDR2, and the LCDR3 comprise the amino acids sequences set forth as SEQ ID NOs: 15, 16, 17, 18, 19 and 20, respectively;   c) the HCDR1, the HCDR2, the HCDR3, the LCDR1, the LCDR2, and the LCDR3 comprise the amino acids sequences set forth as SEQ ID NOs: 25, 26, 27, 28, 29 and 30, respectively;   d) the HCDR1, the HCDR2, the HCDR3, the LCDR1, the LCDR2, and the LCDR3 comprise the amino acids sequences set forth as SEQ ID NOs: 35, 36, 37, 38, 39 and 40, respectively;   e) the HCDR1, the HCDR2, the HCDR3, the LCDR1, the LCDR2, and the LCDR3 comprise the amino acids sequences set forth as SEQ ID NOs: 53, 54, 55, 56, 57, and 58, respectively; or   f) the HCDR1, the HCDR2, the HCDR3, the LCDR1, the LCDR2, and the LCDR3 comprise the amino acids sequences set forth as SEQ ID NOs: 63, 64, 65, 66, 67, and 68, respectively.   
     
     
         3 . The isolated monoclonal antibody or antigen binding fragment of  claim 2 , wherein
 a) the V H  and the V L  comprise the amino acid sequences at least 90% identical to the amino acid sequences set forth as SEQ ID NOs: 1 and 2, respectively;   b) the V H  and the V L  comprise the amino acid sequences at least 90% identical to the amino acid sequences set forth as SEQ ID NOs: 11 and 12, respectively;   c) the V H  and the V L  comprise the amino acid sequences at least 90% identical to the amino acid sequences set forth as SEQ ID NOs: 21 and 22, respectively;   d) the V H  and the V L  comprise the amino acid sequences at least 90% identical to the amino acid sequences set forth as SEQ ID NOs: 31 and 32, respectively;   e) the V H  and the V L  comprise the amino acid sequences at least 90% identical to the amino acid sequences set forth as SEQ ID NOs: 49 and 50, respectively; or   f) the V H  and the V L  comprise the amino acid sequences at least 90% identical to the amino acid sequences set forth as SEQ ID NOs: 59 and 60, respectively.   
     
     
         4 . The isolated monoclonal antibody or antigen binding fragment of  claim 1 , comprising a human framework region. 
     
     
         5 . The isolated monoclonal antibody or antigen binding fragment of  claim 1 , wherein:
 a) the V H  and the V L  comprise the amino acid sequences set forth as SEQ ID NOs: 1 and 2, respectively;   b) the V H  and the V L  comprise the amino acid sequences set forth as SEQ ID NOs: 11 and 12, respectively;   c) the V H  and the V L  comprise the amino acid sequences set forth as SEQ ID NOs: 21 and 22, respectively;   d) the V H  and the V L  comprise the amino acid sequences set forth as SEQ ID NOs: 31 and 32, respectively;   e) the V H  and the V L  comprise the amino acid sequences set forth as SEQ ID NOs: 49 and 50, respectively; or   f) the V H  and the V L  comprise the amino acid sequences set forth as SEQ ID NOs: 59 and 60, respectively.   
     
     
         6 . The isolated monoclonal antibody of  claim 1 , wherein the antibody comprises a human constant domain. 
     
     
         7 . The isolated monoclonal antibody of  claim 1 , wherein the antibody is a human antibody. 
     
     
         8 . The isolated monoclonal antibody of  claim 1 , wherein the antibody is an IgG. 
     
     
         9 . The isolated monoclonal antibody of  claim 1 , comprising a recombinant constant domain comprising a modification that increases the half-life of the antibody. 
     
     
         10 . The isolated monoclonal antibody of  claim 9 , wherein the modification increases binding to the neonatal Fc receptor. 
     
     
         11 . The isolated monoclonal antibody or antigen binding fragment of  claim 1 , wherein the antibody neutralizes group A and group B hMPV. 
     
     
         12 . The isolated monoclonal antibody or antigen binding fragment of  claim 1 , wherein the antibody neutralizes group B2 hMPV. 
     
     
         13 . The isolated monoclonal antigen binding fragment of  claim 1 . 
     
     
         14 . The isolated monoclonal antigen binding fragment of  claim 13 , wherein the antigen binding fragment is a Fv, Fab, F(ab′) 2 , scFV or a scFV 2  fragment. 
     
     
         15 . The isolated monoclonal antibody or antigen binding fragment of  claim 1 , conjugated to an effector molecule or a detectable marker. 
     
     
         16 . A bispecific antibody comprising the monoclonal antibody or antigen binding fragment of  claim 1 . 
     
     
         17 . An isolated nucleic acid molecule encoding the monoclonal antibody or antigen binding fragment of  claim 1 , or a V H  or V L  of the antibody or antigen binding fragment. 
     
     
         18 . The isolated nucleic acid molecule of  claim 17 , comprising
 a) the V H  and the V L  nucleotide sequences set forth as one of SEQ ID NOs: 3 and 4, respectively;   b) the V H  and the V L  nucleotide sequences set forth as one of SEQ ID NOs: 13 and 14, respectively;   c) the V H  and the V L  nucleotide sequences set forth as one of SEQ ID NOs: 23 and 24, respectively;   d) the V H  and the V L  nucleotide sequences set forth as one of SEQ ID NOs: 33 and 34, respectively;   e) the V H  and the V L  nucleotide sequences set forth as one of SEQ ID NOs: 51 and 52, respectively; or   f) the V H  and the V L  nucleotide sequences set forth as one of SEQ ID NOs: 61 and 62, respectively.   
     
     
         19 . The isolated nucleic acid molecule of  claim 17 , wherein the nucleic acid molecule is a cDNA sequence. 
     
     
         20 . The isolated nucleic acid molecule of  claim 17 , operably linked to a promoter. 
     
     
         21 . A vector comprising-a nucleic acid molecule encoding the isolated monoclonal antibody or antigen binding fragment of  claim 1 , or a V H  or V L  of the antibody or antigen binding fragment. 
     
     
         22 . A host cell comprising a nucleic acid molecule encoding the isolated monoclonal antibody or antigen binding fragment of  claim 1 , or a V H  or V L  of the antibody or antigen binding fragment, or a vector comprising the nucleic acid molecule. 
     
     
         23 . A pharmaceutical composition, comprising an effective amount of a monoclonal antibody, an antigen binding fragment thereof, a nucleic acid molecule encoding the monoclonal antibody or antigen binding fragment, or a vector comprising the nucleic acid molecule; and
 a pharmaceutically acceptable carrier,   wherein the monoclonal antibody, or antigen binding fragment thereof, comprise:   a) a heavy chain variable region (V H ) and a light chain variable region (V L ) comprising a heavy chain complementarity determining region (HCDR) 1, a HCDR2, and a HCDR3, and a light chain complementarity determining region (LCDR) 1, a LCDR2, and a LCDR3 of the V H  and V L  set forth as SEQ ID NOs: 1 and 2, respectively;   b) a heavy chain variable region and a light chain variable region comprising aHCDR1, a HCDR2, and a HCDR3, and a LCDR1, a LCDR2, and a LCDR3 of the V H  and V L  set forth as SEQ ID NOs: 11 and 12, respectively;   c) a heavy chain variable region and a light chain variable region comprising HCDR1, a HCDR2, and a HCDR3, and a LCDR1, a LCDR2, and a LCDR3 of the V H  and V L  set forth as SEQ ID NOs: 21 and 22, respectively;   d) a heavy chain variable region and a light chain variable region comprising a HCDR1, a HCDR2, and a HCDR3, and a LCDR1, a LCDR2, and a LCDR3 of the V H  and V L  set forth as SEQ ID NOs: 31 and 32, respectively;   e) a heavy chain variable region and a light chain variable region comprising a HCDR1, a HCDR2, and a HCDR3, and a LCDR1, a LCDR2, and a LCDR3 of the V H  and V L  set forth as SEQ ID NOs: 49 and 50, respectively; or   f) a heavy chain variable region and a light chain variable region comprising a HCDR1, a HCDR2, and a HCDR3, and a LCDR1, a LCDR2, and a LCDR3 of the V H  and V L  set forth as SEQ ID NOs: 59 and 60, respectively,   wherein the monoclonal antibody specifically binds to human metapneumovirus (hMPV) F protein and neutralizes hMPV, and   wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 are identified using Kabat, Chothia or IMGT numbering.   
     
     
         24 . A method of producing an isolated monoclonal antibody, or antigen binding fragment that specifically binds to hMPV F protein, comprising:
 expressing one or more nucleic acid molecules encoding the isolated monoclonal antibody, antigen binding fragment of  claim 1  in a host cell; and   purifying the monoclonal antibody or antigen binding fragment, thereby producing the isolated monoclonal antibody or antigen binding fragment.   
     
     
         25 . A method of detecting the presence of hMPV in a biological sample from a human subject, comprising:
 contacting the biological sample with an effective amount of the isolated monoclonal antibody or antigen binding fragment of  claim 1  under conditions sufficient to form an immune complex; and   detecting the presence of the immune complex in the biological sample, wherein the presence of the immune complex in the biological sample indicates the presence of the hMPV in the sample.   
     
     
         26 . The method of  claim 25 , wherein detecting the detecting the presence of the immune complex in the biological sample indicates that the subject has an hMVP infection. 
     
     
         27 . A method of inhibiting an hMPV infection in a subject, comprising administering an effective amount of pharmaceutical composition of  claim 23  to the subject, wherein the subject has or is at risk of an hMPV infection.